• 제목/요약/키워드: Fluorescence probe

검색결과 274건 처리시간 0.026초

Spectrofluorimetric Determination of Sparfloxacin Using Europium(III) as a Fluorescence Probe in Micellar Medium

  • Kamruzzaman, Mohammad;Alam, Al-Mahmnur;Lee, Sang-Hak;Kim, Young-Ho;Kim, Sung-Hong;Kim, Gyu-Man
    • Bulletin of the Korean Chemical Society
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    • 제33권1호
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    • pp.105-110
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    • 2012
  • A europium (III)-sensitized, spectrofluorimetric (FL) method is presented for the determination of sparfloxacin (SPAR) using an anionic surfactant, sodium dodecyl benzene sulphonate (SDBS). The method is based on the strong fluorescence (FL) enhancement of SPAR after the addition of $Eu^{3+}$ ions as fluorescence probes. The experimental results indicated that the FL intensity of the SPAR-$Eu^{3+}$ system was enhanced markedly by SDBS. The maximum FL emission signal was obtained at about 615 nm when excited at 372 nm. The experimental conditions that affected the FL intensity of the SPAR-$Eu^{3+}$-SDBS system were optimized systematically. The enhanced FL intensity of the system exhibited a good linear relationship with the SPAR concentration over the range of $1.5{\times}10^{-9}-1.2{\times}10^{-7}mol\;L^{-1}$ with a correlation coefficient (r) of 0.9987. The limit of detection ($3{\delta}$) was $4.15{\times}10^{-10}mol\;L^{-1}$ with a relative standard deviation (RSD) of 1.65%. This method was successfully applied for the determination of SPAR in pharmaceuticals, and human serum and urine samples with higher sensitivity, wide dynamic range and better stability. The possible interaction mechanism of the system is also discussed in detail by ultraviolet absorption spectra and FL spectra.

Green Synthesis of Multifunctional Carbon Nanodots and Their Applications as a Smart Nanothermometer and Cr(VI) Ions Sensor

  • Li, Lu;Shao, Congying;Wu, Qian;Wang, Yunjian;Liu, Mingzhu
    • Nano
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    • 제13권12호
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    • pp.1850147.1-1850147.14
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    • 2018
  • In this work, water-soluble and blue-emitting carbon nanodots (CDs) were synthesized from apple peels for the first time via one-step hydrothermal method. The synthetic route is facile, green, economical and viable. The as-prepared CDs were characterized thoroughly by transmission electron microscopy (TEM), X-ray diffraction (XRD), Raman, Fourier transform infrared (FT-IR), X-ray photoelectron (XPS), fluorescence and UV-Vis absorption spectroscopy in terms of their morphology, surface functional groups and optical properties. The results show that these CDs possessed ultrasmall size, good dispersivity, and high tolerance to pH, ionic strength and continuous UV irradiation. Significantly, the CDs had fast and reversible response towards temperature, and the accurate linear relationship between fluorescence intensity and temperature was used to design a novel nanothermometer in a broad temperature range from 5 to $65^{\circ}C$ facilely. In addition, the fluorescence intensity of CDs was observed to be quenched immediately by Cr(VI) ions based on the inner filter effect. A low-cost Cr(VI) ions sensor was proposed employing CDs as fluorescent probe, and it displayed a wide linear range from 0.5 to $200{\mu}M$ with a detection limit of $0.73{\mu}M$. The practicability of the developed Cr(VI) sensor for real water sample assay was also validated with satisfactory recoveries.

Determination of Microviscosity and Location of 1,3-Di(1-pyrenyl) propane in Brain Membranes

  • Kang, Jung-Sook;Kang, In-Goo;Yun, Il
    • Archives of Pharmacal Research
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    • 제20권1호
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    • pp.1-6
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    • 1997
  • We determined the microviscosity of synaptosomal plasma membrane vesicles (SPMV) isolated from bovine cerebral cortex and liposomes of total lipids (SPMTL) and phospholipids (SPMPL) extracted from SPMV. Changes in the microviscosity induced by the range and rate of lateral diffusion were measured by the intramolecular excimerization of 1, 3-di(1-pyrenyl)propane (Py-3-Py). The microviscosity values of the direct probe environment in SPMV, SPMTL and SPMPL were 38.17, 31.11 and 27.64 cP, respectively, at$37^{\circ}C$and the activation energies $(E_a)$ of the excimer formation of Py-3-Py in SPMV, SPMTL and SPMPL were 8.236, 7.448 amd 7.025 kcal/mol, respectively. Probe location was measured by polarity and polarizability parameters of the probe Py-3-Py and probe analogues, pyrene, 1-pyrenenonanol and 1-pyrenemethyl-3${\beta}$-hydroxy-22, 23-bisnor-5-cholenate (PMC), incorporated into membranes or solubilized in reference solvents. There existed a good linear relationship between the first absorption peak of the $^1_a$ band and the polarizability parameter $(n^{2}-1)/(2n^{2}+1)$.The calculated refractive index values for SPMV, SPMTL and SPMPL were close to 1.50, which is higher than that of liquid paraffin (n=l.475). The probe location was also determined by using a polarity parameter $(f-1/2f^{I})$. Here f=$({\varepsilon}-1)/(2{\varepsilon}+1)$ is the dielectric constant function and $f^I=(n^2-1)/(2n^2+1)$ is the refractive index function. A correlation existed between the monomer fluorescence intensity ratio and the solvent polarity parameter. The probes incorporated in SPMV, SPMTL, and SPMPL report a polarity value close to that of 1-hexanol $({\varepsilon}=13.29)$. In conclusion, Py-3-Py is located completely inside the membrane, not in the very hydrophobic core, but displaced toward the polar head groups of phospholipid molecules, e.g., central methylene region of aliphatic chains of phospholipid molecules.

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Membrane-Ordering Effects of Barbiturates on Pure Phospholipid Model Membranes

  • Knag, Jung-Sook;Chung, Young-Za;Cho, Goon-Jae;Byun, Won-Tan;Yun, Il
    • Archives of Pharmacal Research
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    • 제15권3호
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    • pp.196-203
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    • 1992
  • Intramolecular excimer formation of 1, 3-di(1-pyrenyl)propane (Py-3-Py) and fluorescence polarization of 1, 6-diphenyl-1, 3, 5-hexatriene (DPH) were used to investigate the effects of barbiturates on the fluidity of model membranes of phosphatidycholine (SPMVPC), phosphatidylserine (SPMVPS), and phosphatidylinositol (SPMVPI) fractions of synaptosomal plasma membrane vesicles (SPMV) isolated from bovine cerebral cortex. In a dose-dependent manner, barbiturates decreased the excimer to monomer fluorescence intensity ratio (I'/I) of Py-3-Py and increased the anisotropy(r), rotational relaxation time (P), limiting anisotropy $(r_infty)$, and order parameter (S) of DPH in SPMVPC, SPMVPS and SPMVPI. This indicates that barbiturates decreased both the lateral and rotational diffusion of the probes in SPMVPC, SPMVPS and SPMVPI. The relative potencies of barbiturates in ordering the membranes were in the order: pentobarbital > hexobarbital > amobarbital > phenobarbital. This order correlates well with the anesthetic potencies of barbiturates and the potencies for enhancement of $\gamma$-aminobutyric acid-stimulated chloride uptake. Thus, it is strongly suggested that a close relationship might exist between the membrane ordering effects of barbiturates and the chloride fluxes across SPMV.

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Community structure analysis of nitrifying biofilms by 16S rRNA targeted probe and fluorescence in situ hybridization (FISH)

  • 한동우;김동진
    • 한국생물공학회:학술대회논문집
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    • 한국생물공학회 2001년도 추계학술발표대회
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    • pp.282-285
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    • 2001
  • 질산화 생물여과 시스템 내 생물막 안에 존재하는 ammonia oxidizers 및 nitrite oxidizers의 군집 구조 및 공간적 분포를 조사하였다. FISH 분석 결과 생물막 내 숫적으로 우점종을 이루는 미생물은 ammonia oxidizer인 Nitrosomonas spp.로 나타났으며 nitrite oxidizer 인 Nilrospira spp.에 비해 3 ${\sim}$ 5 정도 더 많이 존재하였다. 이는 실협 기간동안 완전한 질산화를 보였지만 반응기가 2 년 이상 nitrite 축적을 위해 높은 free ammonia 농도 빛 낮은 용존 산소 상태에서 운선되어 nitrite oxidizers에 저해를 주었기 때문인 것으로 사료된다. FISH와 결합된 CLSM 관찰 결과 생물막 전체에 걸쳐 ammonia oxidizer가 분포하는 반면 안쪽으로 갈수록 nitrite oxidizers가 분포함을 보였다. 이는 폐수의 ammonium 을 생물막 내 ammon ia oxidizer가 먼저 nitrite로 산화시키고 이를 nitrite oxidizers가 곧바로 nitrate로 산화시키기 때문이다.

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형광제자리부합법을 이용한 치은열구세포 내의 치주염 유발 세균의 관찰 (Visualization of periodontopathic bacteria within crevicular epithelial cells with fluorescence in situ hybridization)

  • 고영경
    • Journal of Periodontal and Implant Science
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    • 제38권4호
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    • pp.691-698
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    • 2008
  • Purpose: Periodontal pathogens can invade the host tissue. Morphologic studies have revealed bacteria within the pocket epithelium, gingival connective tissues, alveolar bone, and oral epithelium. The objective of this study was to visualize and evaluate presence of Porphyromonas gingivalis and Tannerella forsythia in crevicular epithelial cells of periodontally healthy subjects and chronic periodontitis patients. Materials and Methods: A total of 666 crevicular epithelial cells in the samples obtained from 27 chronic periodontitis patients and 9 healthy volunteers were examined. Specific probes for P. gingivalis and T. forsythia and a universal probe for detection of all eubacteria targeting 168 rRNA for fluorescence in situ hybridization was used in conjunction with confocal laser scanning microscopy. Results: 98.99% of sulcular epithelial cells from healthy volunteers and 84.40% of pocket epithelial cells from periodontitis patients were found to harbor bacteria. P. gingivalis and T. forsythia were discovered more often in crevicular epithelial cells from periodontitis patients. Conclusion: P. gingivalis and T. forsythia can invade crevicular epithelial cells and intracellular bacteria may act as a source of bacteria for persistent infection.

Kinetic Mechanism of Nucleotide Binding to Escherichia coli Transcription Termination Factor Rho: Stopped-flow Kinetic Studies Using ATP and Fluorescent ATP Analogues

  • Kim, Dong-Eun
    • Biotechnology and Bioprocess Engineering:BBE
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    • 제9권1호
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    • pp.23-34
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    • 2004
  • Escherichia coli transcription termination factor Rho catalyzes the unwinding of RNA/DNA duplex in reactions that are coupled to ATP binding and hydrolysis. Fluorescence stopped-flow methods using ATP and the fluorescent 2'(3')-O-( N-methylanthraniloyl) derivatives (mant-derivatives) of ATP and ADP were used to probe the kinetics of nucleotide binding to and dissociation from the Rho-RNA complex. Presteady state nucleotide binding kinetics provides evidence for the presence of negative cooperativity in nucleotide binding among the multiple nucleotide binding sites on Rho hexamer. The binding of the first nucleotide to the Rho-RNA complex occurs at a bimolecular rate of 3.6${\times}$10$\^$6/ M$\^$-1/ sec$\^$-1/ whereas the second nucleotide binds at a slower rate of 4.7${\times}$10$\^$5/ M$\^$-1/ sec$\^$-1/ at 18$^{\circ}C$, RNA complexed with Rho affects the kinetics of nucleotide interaction with the active sites through conformational changes to the Rho hexamer, allowing the incoming nucleotide to be more accessible to the sites. Adenine nucleotide binding and dissociation is more favorable when RNA is bound to Rho, whereas ATP binding and dissociation step in the absence of RNA occurs significantly slower, at a rate ∼70- and ∼40-fold slower than those observed with the Rho-RNA complex, respectively.

VisiSens 산소 평면광 센서 시스템을 이용한 식물 잎 표면의 산소분포 가시화 (Visualization of oxygen distribution on leaf surfaces using VisiSens oxygen planar optode system)

  • 황배근;김혜정;이상준
    • 한국가시화정보학회지
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    • 제14권1호
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    • pp.51-56
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    • 2016
  • Oxygen is a key factor in aerobic reactions and most biological activities. Visualization of oxygen distribution of a chemical process or biological system has been a very challenging object despite of its significance and potential impact. To monitor and visualize the spatial distribution of oxygen concentration, various techniques such as electro-chemical probe, polarographic electrode, LIF(laser-induced fluorescence) and so on have been introduced. Oxygen planar optode which utilizes the oxygen quenching of fluorescence light is one of the currently available methods for time-resolved visualization of oxygen distribution on a planar surface. In this study, we utilized VisiSens oxygen planar optode system to visualize the spatial distribution of oxygen concentration on leaves of Korean azalea. As a result, temporal variation of oxygen concentration distribution caused by respiratory activity of the leaf could be quantitatively monitored.

Use of Ratiometric Probes with a Spectrofluorometer for Bacterial Viability Measurement

  • Cleach, Jerome;Watier, Denis;Le Fur, Bruno;Brauge, Thomas;Duflos, Guillaume;Grard, Thierry;Lencel, Philippe
    • Journal of Microbiology and Biotechnology
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    • 제28권11호
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    • pp.1782-1790
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    • 2018
  • Assessment of microorganism viability is useful in many industrial fields. A large number of methods associated with the use of fluorescent probes have been developed, including fluorimetry, fluorescence microscopy, and cytometry. In this study, a microvolume spectrofluorometer was used to measure the membrane potential variations of Escherichia coli. In order to estimate the sensitivity of the device, the membrane potential of E. coli was artificially disrupted using an ionophore agent: carbonyl cyanide 3-chlorophenylhydrazone. The membrane potential was evaluated using two ratiometric methods: a Rhodamine 123/4',6-diamidino-2-phenylindole combination and a JC-10 ratiometric probe. These methods were used to study the impact of freezing on E. coli, and were compared with the conventional enumeration method. The results showed that it was beneficial to use this compact, easy-to-use, and inexpensive spectrofluorometer to assess the viability of bacterial cells via their membrane potential.

자성 산화철(iron oxide) 나노입자를 이용한 DNA 센서 개발 (Development of DNA Sensor Using Magnetic Iron Oxide Nanoparticle)

  • 남기창;송광섭
    • 전자공학회논문지SC
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    • 제48권6호
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    • pp.51-56
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    • 2011
  • 자성 산화철 나노입자(iron oxide nanoparticle, ${\gamma}-Fe_2O_3$) 표면을 기능성 유기 분자를 이용하여 아민기($-NH_2$), 카르복실기(-COOH)로 표면 처리 하였으며, 이들 기능기로 표면 처리된 산화철 나노입자를 FT-IR을 이용하여 나노입자 표면을 분석하였다. 아민기, 카르복실기로 표면처리된 산화철 나노입자 표면에 특정 배열을 갖는 21-base pair 길이의 프로브 DNA를 고정하였고, 형광 라벨(Cy5)이 부착된 상보적, 비상보적 타게트 DNA를 이용하여 고정된 프로브 DNA와 hybridization을 진행하였다. 각각의 상보적, 비상보적 타게트 DNA와 hybridization 처리한 산화철 나노입자를 confocal microscopy를 이용하여 관찰하였으며, 그 결과 산화철 나노입자를 이용하여 특정 배열의 DNA검출에 성공하였다.